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Everett Price
Everett Price

INTACT - 3 Lei

Development in mass spectrometry (MS) and software has enabled the characterization and quantification of intact glycopeptides in complex biological matrixes. Zhu et al. have identified and quantified N-glycopeptides of Hp based on electron-transfer higher-energy collision dissociation (ET-hcD), MS/MS fragmentation, and Byonic software (Zhu et al., 2019b). We have previously used a glycopeptide method based on 18O/16O C-terminal labeling and multiple reaction monitoring (MRM) to quantify N-glycopeptides of the 40-kDa band in liver diseases (Zhang et al., 2019).

INTACT - 3 lei

As shown in Figure 3A, 26 intact O-glycopeptides on four O-glycosylation sites (Thr126, 317, 323, and Ser316) were identified on the Hp protein and corresponded to 18 types of glycan compositions. Most of them were elevated in HCC as compared to LC. As shown in Supplementary Table S1, 57.69% of the changed glycopeptides were observed in seven patients and 96.15% in at least four patients. In addition, the majority of the glycoforms were located on Ser316 and Thr317, while one glycoform was located on Thr126 and Thr323. Moreover, one glycoform (H1N4) on Thr317 could be detected in HCC; however, this was absent in cirrhosis. Among these O-glycopeptides on Hp, the intensity of HYEGS316TVPEK (H1N1S1) was the highest, and it was significantly increased in HCC patients (Figure 3B, p

MRM for validation. (A) MRM transitions for monitoring the O-glycopeptide and the unique peptide of Hp. (B) HYEGS316TVPEK (H1N1S1) of Hp from 12 LC patients and 12 HCC patients was detected, and the difference was statistically significant. (C) O-glycopeptide abundance was divided by the unique peptide abundance to separate out the contribution of protein concentration. The result showed that the increased HYEGS316TVEPK (H1N1S1) was caused by elevated protein expression in HCC. LC-IG, the intact O-glycopeptide in LC; HCC-IG, the intact O-glycopeptide in HCC; LC-IG/Pro, the intact O-glycopeptide abundance divided by the protein abundance in LC; HCC-IG/Pro, the intact O-glycopeptide abundance divided by the protein abundance in HCC.

Figure 1. The procedural framework of the 4-plex ddPCR for specific detection of intact V. parahaemolyticus cells. The bacterial solution containing intact cells were added to the PCR reaction. The cells were dispersed into droplets prior to PCR-amplification. The targets of tlh (green), tdh (red), ureR (yellow), and orf8 (purple) were determined based on fluorescent probes.

Methods: Multivariate linear models analyzed the available data on CSF biomarkers and frequency of green tea consumption of 722 cognitively intact participants from the Chinese Alzheimer's Biomarker and LifestylE (CABLE) database, and we additionally detected the interaction effects of tea consumption with APOEɛ4 status and gender using a two-way analysis of covariance.

Our objective was to analyze the spatial patterns and structural connectivity of intact and primary forests in northern Sweden with focus on the Scandinavian Mountain region; one of the few remaining large European intact forest landscapes.

We revealed a contiguous, connected and semi-connected intact forest landscape forming a distinct Green Belt south to north along the mountain range. Almost 60% of the forestland remains intact, including contiguous clusters 10,000 ha and larger. The connectivity is particularly high in protected areas with primary forests outside contributing substantially to overall connectivity. We found gaps in connectivity in the southern parts, and furthermore low or absent connectivity across the whole inland and coastal areas of northern Sweden.

The concept of green infrastructure, which as a fervent EU initiative is integrated into one or more policy sectors in all member states (Slätmo et al. 2019), aims to secure biodiversity, habitat resilience and ecosystem services at multiple spatial scales (Liquete et al. 2015). Thus, green infrastructure promotes landscape-scale and holistic planning based on known conservation core areas and their functional connectivity in the existing matrix, as, for example, the Yellowstone to Yukon Conservation Initiative in North America (Mahr 2007) and the Australian Alps to Atherton Connectivity Conservation Area (Pulsford et al 2010). In practice this implies that green infrastructure requires a spatio-temporal perspective ranging from local species occurrence and microsites, to habitats, landscapes and entire regions (e.g., Heller and Zavaleta 2009; Gustafsson et al. 2012). Furthermore, a functional green infrastructure intrinsically relies on a continuum of intra- and inter-connected land cover types and their transitions, temporal land-use changes, natural succession and dynamics, and also restoration of transformed matrix surrounding the core areas (Sayer 2009; Chazdon et al. 2016; Chazdon 2018). Hence, for green infrastructure focusing on forest ecosystems, intact forest landscapes play a paramount role. The landscape-scale and holistic approach in the green infrastructure concept is recognized in the Aichi target 7 on sustainable management, biodiversity and conservation in target 11 on setting aside a minimum of 17% of terrestrial areas, and in target 15 on restoring degraded ecosystems (CBD 2010). Although frequently promoted, however, landscape approaches aiming for advanced nature conservation are often not successful in practice and hence need to be further developed (e.g., Chazdon et al. 2017; IPBES 2018).

There were important differences among the mountain SRs. The north mountain SR had the overall highest connectivity, i.e. the largest areas with pCF-clusters of any connectivity class (Fig. 4; Table 3) and also a relatively even distribution of clusters across all connectivity classes. Both the central and south mountain SRs had areas with the highest connectivity class weighted to total forest land (21.8% and 17.8%, respectively) at comparable proportion to the north mountain SR (19.6%). However, the weighted areas of low, moderate and high connectivity classes, especially the two latter, were much smaller which thus shows a skewed connectivity distribution. For the south mountain SR in particular, the overall connectivity within intact forest landscapes relies on the fraction of highest connectivity.

The forest landscapes of the European north are dominated by systematic clear cutting rotation forestry and thus have undergone extensive transformation (e.g. Peura et al. 2018). Data that allows change detection of how, where and to what extent this transformation have influenced the natural landscape configuration, such as the data applied in this study, are very valuable. Detailed spatial analyses of remaining intact forest landscapes and primary forests on local and regional scale, clearly fill a knowledge gap for national (e.g., Angelstam et al. 2020) and European and other pan-national contexts (e.g., Sabatini et al 2018), beyond adding functional dimensions to green infrastructure planning. Similar approaches in other European regions, and potentially also for other land-cover types than forests, would provide a much needed supra-national perspective on functional green infrastructure assessments (Slätmo et al. 2019; Hermoso et al. 2020).

For the mountain and northernmost parts of the study region, a proportion of the intact forest consists of subalpine mountain birch. Notably, the density analysis step-wise omitted smaller, isolated and elongated forests and woodlands in high altitude mountain valleys dominated by mountain birch. Thus, the main connectivity route was concentrated to larger, more contiguous, and more coniferous dominated forests. The highest connectivity was detected in the central mountain SR where the proportion of mountain birch forest was estimated to be the lowest. This partly indicates that connectivity there largely is associated with coniferous forests but also is the effect of our assumptions for a connectivity analysis (i.e. that only larger contiguous forest clusters were taken into account) and our definition of an intact forest landscape cluster (see the methods section). By contrast, the connectivity in the north mountain SR is associated with a higher share of mountain birch forests, in a landscape characterized by large mires, rocky outcrops, etc., land without or with low cover of trees (SLU 2018). This mosaic landscape naturally implies a lower level of natural forest connectivity, as detected in this study. However, it is also assumed that limited accessibility for forestry and already large protected areas explains the patterns in the north SR. Thus, our connectivity analysis is found to be sensitive to differences in natural landscape configuration. However, it should be stressed that open and semi-open forests on both productive and non-productive forest land can harbor significant continuity values (Hemäläinen et al. 2017) and hence contribute to functional green infrastructure.

There is an increasing interest in identification of remaining intact and wilderness forest areas on pan-national and global scales, and there is a critical need for policy recognition and protection of such areas. Recent publications include for example Haddad et al. (2015), Heino et al. (2015), Potapov et al. (2017), Jones et al. (2018), Müller et al. (2018) and Watson et al. (2018). Our study contributes precision and resolution in one of those areas that have been identified, at a scale that allows for strategic and operational planning. In addition to the existing protected areas, we have identified forest areas for future conservation, restoration and adaptive management in a structured, systematic manner across a large region. Our analysis revealed general pattern of structural connectivity of primary forests in intact forest landscapes, however, without distinguishing between different forest types and assuming homogeneous non-forest matrix between those areas. Further studies are needed for understanding the functional ecological aspects of intact forest landscapes and connectivity from the perspective of different groups of organisms in boreal forest landscapes, both forest specialists and generalists, especially taking into account their ecological traits, habitat and landscape requirements and sensitivity to forest clear cutting and other anthropogenic disturbances. Also, further studies on natural (e.g. topography, spatial distribution of open mires and major waterbodies) and human (e.g. historic land-use, transport infrastructure) causes to the patterns of remaining primary forests and intact forest landscapes needs further attention (Axelsson and Östlund 2001; Mikusiński et al. 2003; Angelstam et al. 2004; Boucher et al. 2009). 041b061a72




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